Fret Imaging of Neuronal Contact with Diamond Nanopillar Electric Field Sensors
il y a 2 semaines
**FRET imaging of neuronal contact with diamond nanopillar electric field sensors**:
- Réf **ABG-127591**
- Stage master 2 / Ingénieur
- Durée 6 mois
- Salaire net mensuel ~600€
- 16/12/2024
- Université Paris Saclay
- Lieu de travail- Gif-Sur-Yvette Ile-de-France France
- Champs scientifiques- Sciences de l’ingénieur
- Biotechnologie
- Santé, médecine humaine, vétérinaire
- Mots clés- Nanosensors, biophysics, fluorescence imaging, diamond nanopillars, electric field sensors- 31/01/2026
**Établissement recruteur**:
**LuMIn** team of optics physicists has a long expertise of translating tools and methods of nanosciences and microscopies to biomedical research. Being also trained in quantum physics, it now applies quantum sensing to life science. The **I2BC**team has expertise in biochemistry and biophysics. Its research focuses on nanodetection of biomolecular interactions using FRET, allowing to measure membrane-protein distances with sub-nanometric resolution. The two teams obtained preliminary results together.
**Description**:
Neurons communicate with each other via synapses. Little is known about electrical potential at the synapse submicron scale, even though it plays a key role in synaptic transmission. LuMIn team is setting up an optical sensor of neuron electrical activity with 200 nm and 1 ms spatial and temporal resolutions, based on quantum properties of nitrogen-vacancy (NV) defects in diamond.
- As electric field **E** can be screened by culture medium ions on distance as short as 1 nm (Debye layer), it is critical that neuronal branches are tightly connected to the pillars, as it seems to be the case based on our preliminary electron microscopy images. However, to ascertain a tight contact at the macromolecular level, further studies are necessary.
- We propose to use **FRET** (Förster resonance energy transfer) **[2]** to estimate the distance between the neuron membrane and the pillar, the effect taking place on distance of the order of a few nanometers only. The FRET pair donor will be semiconductor nanocrystals (QDots), while the acceptor will be a fluorogenic cell membrane dye. QDots will be conjugated to laminin used as a coating onto the pillar to promote neuron adhesion.
- We will start with an _in vitro_ proof of principle using **Giant Unilamellar Vesicle** (GUV, produced by **I2BC** team), mimicking neuron membrane. The intern will be in charge of (i) Qdots conjugation to laminin and the subsequent pillar array coating, (ii) the preparation of GUV synthetic model and their membrane labeling with dyes (MemBright), and (iii) FRET imaging of GUV on QDots coated nanopillar using fluorescence confocal microscopy. Once our model strategy validated, we will repeat the experiment with **mouse primary neurons**cultured by **LuMIn** team onto the nanopillar array.
- 1. L. Hanlon et al., _Neurophotonics_ **7**, 035002 (2020)
- 3. Haziza, S. [ ], Treussart*, F, Simonneau*, M, _Nat Nanotechnol_ **12**, 322 (2017).
**Profil**:
This interdisciplinary project will mobilise ground-breaking imaging techniques such as:
- Förster resonance energy transfer (FRET)
- electric field sensors
- Confocal microscopy
As well as experimental techniques such as:
- cell culture
- In vitro biomimetic systems (supported lipid bilayers)
**Prise de fonction**:
- 20/01/2025
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